Correlation of multidrug resistance (MDR1) protein expression with functional dye/drug efflux in acute myeloid leukemia by multiparameter flow cytometry: identification of discordant MDR-/efflux+ and MDR1+/efflux- cases

Abstract

Resistance to chemotherapy is a major factor limiting successful treatment of acute myeloid leukemia (AML); one of the best characterized drug resistance mechanisms is extrusion of drugs by the energy-dependent multidrug resistance (MDR1) transport protein. Expression of MDR1 is common in AML and has been linked to lower remission induction rates and decreased remission durations. Because MDR1 efflux function may be modified by drugs such as cyclosporin A, accurate identification of MDR1+/efflux+ AML cases will be critical to identify patients who may benefit from therapies that contain such MDR1 modulators. We have optimized single and multiparameter flow cytometric assays to detect efflux of drugs or fluorescent dyes by previously cryopreserved AML blasts. These assays allowed precise identification of efflux by leukemic blasts, and correlation with CD34 and MDR1 expression. We subsequently studied a series of 60 previously untreated AML cases. Functional efflux was identified in 39 cases and was significantly correlated with MDR1 expression (P = .0002). However, discrepant cases were identified; 10 cases were efflux+ without significant MDR1 expression, whereas 6 MDR1+ cases were efflux-. There was also a highly significant correlation of efflux with CD34; 31 (79%) of the 39 efflux+ cases were CD34+ in comparison with only 5 (24%) of the 21 efflux- cases (P < .0001). Multivariate analysis showed that efflux was significantly associated with independent effects of both CD34 (P = .0011) and MDR1 expression (P = .034); the majority of efflux+ cases were CD34+, whereas 5 of the 6 MDR1+ efflux- cases lacked CD34 expression. Cyclosporin A blocked efflux in all but 2 cases regardless of MDR1 expression. Functional efflux in AML is frequently detected without the classic MDR1+ phenotype indicating that alternate non-MDR1-mediated efflux mechanisms may be important. Efflux assays may better identify patients who would benefit from therapies that include efflux modulators.

P‐glycoprotein and multidrug resistance‐associated protein, but not lung resistance protein, lower the intracellular daunorubicin accumulation in acute myeloid leukaemic cells

“…activity but low or no Pgp expression. Similar ®ndings have been reported previously (Lamy et al, 1995;Leith et al, 1995).…”

MDR 1 expression is an independent prognostic factor for response and survival in de novo acute myeloid leukaemia

“…In untreated de novo AML patients MDR 1 is expressed in 19-75% of the patients (Campos et al, 1992;Leith et al, 1995;Boekhorst et al, 1995Boekhorst et al, , 1993Holmes et al, 1994;Guerci et al, 1995). In these patients, MDR 1 is frequently associated with an immature phenotype (CD34) and autonomous AML growth in vitro (Boekhorst et al, 1995).…”

“…Expression of transmembrane transporter proteins in tumour cells has been identified as an important cause of chemotherapy resistance (Bradley et al, 1988;Arceci, 1993). Of these, the Multidrug Resistance (MDR 1) gene encodes a membrane P-glycoprotein (P-gp), which is expressed in 19-75% of untreated AML and is associated with a low probability of achieving a complete remission (CR) (Campos et al, 1992;Leith et al, 1995;Boekhorst et al, 1995;Boekhorst et al, 1993).…”

Transport proteins in drug resistance: detection and prognostic significance in acute myeloid leukaemia

“…even suboptimal methods. Those highest Pgp expressers have an increase of approximately 30-50% of steady-state daunomycin accumulation after blocking its efflux with PSC-833 [12][13][14][15].…”